Biopesticide composition and process for controlling insect pests

ABSTRACT

The subject invention concerns a novel anal useful biopesticide with activity against insect pests such as boll weevil, sweet potato whitefly and cotton fleahopper. The biopesticide of the subject invention comprises an entomopathogenic fungus having virulence against a targeted insect pest(s), an arrestant anal feeding stimulant for the targeted insect pest(s) anal, optionally, a pheromone for the targeted insect pest(s). A preferred fungus is a Beauveria bassiana, preferably Beauveria bassiana, ATCC-74040 (ARSEF-3097). By using this novel fungus, or mutant thereof, in the composition of the present invention, Boll weevils, sweet potato whiteflies anal cotton fleahoppers can be controlled without environmental and public safety hazards presented by chemical control agents.

This application is a continuation of application Ser. No. 07/892,488filed Jun. 3, 1992 which is a continuation of application Ser. No.07/638,489, filed Jan. 9, 1991, now abandoned.

FIELD OF THE INVENTION

This invention relates, in general, to insecticides for controllinginsect pests and methods for using such compositions. In one aspect,this invention relates to a biopesticide composition for controllinginsect pests, wherein the composition comprises an entomopathogenicfungus having virulence against the insect pest(s), an arrestant andfeeding stimulant for the insect pest(s), and, optionally, a pheromonefor the insect pest(s). In a further aspect, this invention is directedto the use of such compositions. In another aspect, this inventionrelates to an isolate of Beauveria bassiana which, when in its pureform, has virulence against boll weevils, sweet potato whiteflies andcotton fleahoppers characteristic of Beauveria bassiana culture depositATCC-74040 (ARSEF 3097).

BACKGROUND OF THE INVENTION

The cotton boll weevil is easily one of the most notorious agriculturalpests in the world. It occurs in all principal cotton-growing areas ofCentral and South America and the United States except parts ofCalifornia. Wherever it is present, it is the key pest in cotton. Theinjury is caused by adults and the larvae or grubs. The adult weevilschew into or puncture the squares and bolls, and with their long slenderbills, feed on the inner tissues. The eggs are laid in these holes, andthe hatching grubs bore into the boll or square, causing the squares todrop off or wither and dry on the plant. This feeding either results indirect destruction of the flower or a reduction of fiber content in theboll. Losses can be so great as to be limiting. In 1982, damage tocotton in the United States was estimated at $429 million dollars. Thisfigure has and is expected to continue to increase.

Chemical insecticides and cultural eontrois are currently employed inthe control of boll weevil. These have associated problems and are notcompletely effective. There is a definite need for alternative materialsthat can be used in a complementary fashion with existing controls andto replace control agents that may lose efficacy due to resistance orother factors.

Management strategies for boll weevil control seek to attack the insectat the weakest point in its life cycle. Thus, early season adult controlis based on insecticide treatment at pinhead square formation to killoverwintering adults prior to their damaging the current crop andbeginning reproduction. Because insecticides kill benefieials and pestsalike, the predator-prey balance is broken. Surviving insect pestsproliferate due to the decreased populations of beneficial predatoryinsects, and additional insecticide applications become necessary forcontrol of later pests, particularly the bollworm complex, Heliothisspp. Thus, removal of the early season boll weevil is tantamount forproduction of a viable lint crop. With increased emphasis on protectionof the environment, reduction in the use of pesticides, and withincreased urbanization and awareness, the need for biopestieides andinsect management strategies is acute, and it should become an importantfactor in sustainable agricultural concepts now being rigorouslysupported by state and federal institutions. One of the most successfulmaterials, Bacillus thuringiensis, has been tested extensively against aboll weevil but remains ineffective.

The sweet potato whitefly Bemisia tabaei (Gennadius) has appeared onpoinsettias in California, Florida, Georgia and North Carolina. During1981, the sweet potato whitefly was responsible for crop and marketlosses of 100 million dollars in cotton, eueurbits and lettuce inCalifornia and Arizona. The whitefly is increasingly a problem inFlorida where, in 1986, this whitefly caused approximately 2 milliondollars of damage to Florida's 8-10 million dollar poinsettia crop.

The sweet potato whitefly is also a pest of international importance,having been found on host plants throughout the mideast Caribbean andCentral America. This insect is now known to feed on more than 500different plants, many of which are of importance in the Caribbean andFlorida. For example, cassava, sweet potato, squash, tomato, beans,lettuce, cotton, pepper, carrot, cucumber, eggplant, and water melon areall known hosts. This species of whitefly severely impacts infestedplants by its feeding, production of honeydew with resultant growth ofsooty mold, and transmission of plant pathogens. Most extensive lossesto this pest have been through direct feeding damage and indirect damagethrough transmission of plant diseases.

Whitefly-borne diseases are of major importance in tropical andsubtropical agriculture. More than 70 diseases caused by viruses andmicroorganisms are known to be transmitted by whiteflies, with most ofthem being transmitted by the sweet potato whitefly. In Puerto Rico,this whitefly is a vector of at least seven diseases. One of thesediseases is the bean golden mosaic virus, a disease affecting manylegumes.

The sweet potato whitefly has proven to be very difficult to controlwith conventional pesticide applications. Many factors contribute to thelack of control obtained with pesticides. The most important factor isthat this whitefly has demonstrated a broad spectrum of resistance tochlorinated hydrocarbons, organophosphorus, carbamate, and syntheticpyrethroid insecticides. Very few commercially available pesticides areeffective against whiteflies, and those which do work are only effectiveif care is taken to make a very thorough application of the insecticideseveral times a week. The sweet potato whitefly spends most of its lifeon the undersides of leaves, therefore, growers must adjust theirmanagement practice to permit increased pesticide coverage there. Thespacing of the plants must be such that the chemical spray can penetratethe canopy and reach all surfaces of the plants.

In addition to being largely ineffective, and difficult and costly toapply, chemical control of these pests has other significant drawbacks.For example, the use of chemical pesticides presents the furtherdisadvantages of polluting the environment, creating potential healthhazards to agricultural workers and to consumers, development ofresistance to chemicals in target pest species, detrimental effect ofthese chemicals on non-target species resulting in secondary pestoutbreaks, and phytotoxic reaction by treated plants.

Because of the problems associated with the use of chemical pesticides,safer and more effective methods of control for insect pests are clearlyneeded. Although biological control agents are actively being soughtafter, to date no biological control agent has been commerciallysuccessful for the control of the boll weevil and the sweet potatowhitefly.

Biological control agents have been tried; however, availability,limited host range, cost and reliability have reduced the potential forimplementing the use of these biological control agents. The developmentof a broad spectrum of pesticides would reduce the need for many of thepetrochemically based pesticides. By using fungi to control insectpests, the potential for resistance development is minimized, which, inturn, will stabilize many of the pest management programs.

In many instances, fungi used to control insect pests have not had theeffectiveness required for commercial use. Therefore, there also existsa need for enhancing the effectiveness of such entomopathogenic fungi.

SUMMARY OF THE INVENTION

Accordingly, there is provided a biopesticide composition forcontrolling targeted insect pests, for example, the boll weevil, cottonfleahopper and the sweet potato whitefly. The biopesticide compositioncomprises an entomopathogenic fungus having virulence against thetargeted insect pest(s), an arrestant and feeding stimulant, and,optionally, a pheromone. The entomopathogenie fungus may be a Beauveriabassiana, preferably Beauveria bassiana culture deposit ATCC-74040(ARSEF 3097), and routants thereof which substantially retain thevirulence of the parent strain against boll weevils, sweet potatowhiteflies and cotton fleahoppers. The fungus may also be Paecilomycesfumosoroseus, designated Apopka, culture deposit ATCC 20874, and mutantsthereof which substantially retain the virulence of the parent strainagainst whiteflies.

The arrestant and feeding stimulant is preferably one derived fromcotton-plant parts and square components, preferably from the cottonseedand/or the water extract of the calyx and androecium parts of the cottonplant.

The pheromone is preferably a sex and/or aggregating attractant specificto the insect pest(s) of interest.

DETAILED DESCRIPTION OF THE INVENTION

A biologically pure culture of a novel isolate of Beauveria bassiana ofthe subject invention has been deposited in the American Type CultureCollection (ATCC), 12301 Parklawn Drive, Rockville, Md. 20852 andarranged accession No. ATCC 74040. This same novel isolate was alsoaccessioned into the USDA-ARS Collection of Entomopathogenic FungalCultures as ARSEF 3097 at which time the culture was confirmed to be aBeauveria bassiana (Balsamo) Vuillemin. The USDA-ARS accession recordfor this strain has been marked for restricted distribution, whereby theUSDA-ARS will not release this fungus for use by any laboratory withoutpermission of the depositor.

    ______________________________________                                        Culture       Accession No.                                                                             Deposit Date                                        ______________________________________                                        Beauveria bassiana                                                                          ATCC-74040  March 11, 1991                                      ______________________________________                                    

As noted above, the USDA-ARS deposit is restricted and will not releasedwithout permission of the depositor. The subject culture is depositedunder conditions that assure that access to the culture will beavailable during the pendency of the patent application to onedetermined by the Commissioner of Patents and Trademarks to be entitledthereto under 37 C.F.R. § 1.14 and 35 U.S.C. § 1.22. The ATCC depositwill be available as required by foreign patent laws in countrieswherein counterparts of the subject application, or its progeny, arefiled. However, it should be understood that the availability of adeposit does not constitute a license to practice the subject inventionin derogation of patent rights granted by governmental action.

Furthermore, the subject culture deposit will be stored and madeavailable to the public in accord with the provisions of the BudapestTreaty for the Deposit of Microorganisms, i.e., it will be stored withall the care necessary to keep it viable and uncontaminated for a periodof at least five years after the most recent request for the furnishingof a sample of the deposit, or in any case, for a period of at leastthirty (30) years after the date of deposit or for the enforceable lifeof any patent which may issue disclosing the culture. The depositoracknowledges the duty to furnish a sample when requested, due to thecondition of the deposit. All restrictions on the availability to thepublic of the subject culture deposit will be irrevocably removed uponthe granting of a patent disclosing it.

The entomopathogenic fungus Beauveria bassiana is an imperfect fungus(Fungi Imperfecti) and the subdivision Deuteromycotonia. The genusBeauveria Vuill is within the Class Deuteromycetes and is distinguishedfrom other genera by having conidia that are borne singly, notcatenulate and having the fertile portion of the conidiophore zig-zag inshape and drawn out at the tip. The species Beauveria bassiana hasspherical not ellipsoid conidia measuring 2-3 micrometers by 2-2.5micrometers and with conidiophores forming dense bunches. The novelisolate of Beauveria bassiana is the first known fungus of this specieswhich is highly virulent to boll weevils, sweet potato whiteflies andcotton fleahoppers.

Like most entomogenous fungi, Beauveria bassiana initiates infection bya germinating spore (conidium) attaching to and subsequently penetratingthe cuticle of the insect host. The claimed Beauveria bassiana attachesvery securely to the cuticle of the targeted insect pest and istypically not removed by the grooming activities thereof. This mayaccount somewhat for the high virulence of the fungus. As the funguspenetrates the target pest cuticle, the invasive hyphae begin to enterthe host tissues and ramify through the hemocoel. Hyphal bodies orsegments of the hyphae distribute throughout the hemocoel, filling thedying insect with mycelium. Emergence hyphae grow out through theinsect's integument and produce spores on the external surface of thehost. These spores, or conidia, are dispersed and capable of infectingnew host insect pests.

The fungus works rapidly; a remarkable 80 to 90 percent kill of theadult boll weevil occurs within 3 to 10 days of application.Significantly, more than 90 percent of the kill occurred within one weekof application. Spore concentrations of from about 2×10⁸ to about 2×10¹⁴spores per milliliter of carrier can be used. Spore application rates offrom about 1×10¹² to about 10×10¹² per acre can be used, preferablyabout 4.5×10¹² to about 6.25×10¹² conidia per acre.

This particular Beauveria bassiana may be cultured and mass produced bymethods used to culture Beauveria spp. See for example, U.S. Pat. No.4,925,663; Micobial Control of Pests and Plant Diseases 1970-1980,published by Academic Press, pages 471-473 (1981; edited by H. D.Burges); and Feng et al., J. Invertebrate Pathology, Vol. 46, no. 3,November 1985, page 260, the disclosures of which are herebyincorporated by reference.

In another aspect of the present invention, a biopesticide compositionfor controlling targeted insect pests is provided. The biopesticidecomposition comprises an entomopathogenic fungus having virulenceagainst the targeted insect pests, an arrestant and feeding stimulant,and, optionally, a pheromone. In this manner, contact with the fungus isenhanced by the attractive properties of the arrestant and feedingstimulant and, optionally, in conjunction with a sex or aggregatingpheromone. As noted above, in the discussion of the Beauveria bassiana,the mode of infection of fungi is generally by cuticular penetration bythe germ tube of the fungal conidia and may also occur through therespiratory or alimentary tract (such as mouth parts). Additionally,ingestive fungal spores voided in the feces may provide another sourceof contact with the cuticle of the targeted insect pest. Death in thehost may occur either by release of fungal toxins or by tissuedestruction. The fungal growth range is between 40° and 95° F. in a widerange of humidity with high humidity necessary to germinate spores andto increase spore production on the crops or plants being protected bythe biopesticide.

Entomopathogenic fungi which may be used in the biopesticide compositionof the present invention include but are not limited to:

Beauveria spp.

Metarhizium spp.

Paecilomyeus spp.

Akanthomyees spp.

Gibellula spp.

Pseudogibellula spp.

Hymenostilbe spp.

Spicaria spp.

Nomuraea spp.

Cephalosporium spp.

Sporotriehum spp.

Torubiella spp.

Cordyceps spp.

Aspergillus spp.

Entomopathora spp.

Paeeilomyees spp.

The following fungi are specific examples which are suitable in thebiopesticide composition of the present invention. Beauveria bassianaNo. RS-252 available from Abbott Laboratories, Chicago, Ill., which isuseful against cotton boll weevils.

Beauveria bassiana culture deposit ATCC-74040 (also deposited with theUSDA-ARS having accession No. ARSEF-3097) useful in the control ofcotton boll weevil, sweet potato whitefly, and cotton fleahopper.

Paecilomyees fumosoroseus, designated Apopka, culture deposit ATCC-20874(See U.S. Pat. No. 4,942,030, the disclosure of which is incorporatedherein by reference) useful in the control of the sweet potato whitefly,onion thrips, beet army worm, Colorado potato beetle, gypsy moth,two-spotted spider mite, flower thrips, Citrus mealybug, and Solanummealybug.

Beauveria bassiana No. 447 culture deposit ATCC-20872 (See U.S. Pat. No.4,925,663, the disclosure of which is incorporated herein by reference)useful in the control of imported fire ants.

The arrestant and feeding stimulant may be broad based to a variety ofinsect pests or tailored to a specific insect pests. In regards toinsect pests of cotton, the arrestant and feeding stimulant ispreferably one derived from cotton plant parts and square components,preferably from cottonseed and/or the water extract of the calyx andandroecium parts of the cotton plant. Examples of such insect pestsinclude the boll weevil, sweet potato whitefly, and the cottonfleahopper.

A broad based feeding stimulant suitable for use in the biopesticidecomposition of the present invention is a combination of protein,carbohydrates and lipid oil which may be formulated as an aqueoussuspension, wettable powder, dry flowable water, dispersible granule,granular bait or dust. These three ingredients may be present atconcentrations of 10 to 100 percent by weight of the feeding stimulant.The ratio of the protein, carbohydrate and lipid oil components ispreferably in the range of about 1:1:0.2 to about 1:2:0.03 with aparticularly preferred ratio being 42.5:55:2.5. The protein sources arepreferably high protein sources having about 40 to about 70 percent byweight protein. Furthermore, vegetable protein is preferred over animalprotein. Examples of vegetable protein is the protein included insoybeans, cotton seed and the like. Particularly preferred is cottonseedflour which has been defatted (preferably less than 4% by weight fat),low gossypol (preferably less than 0.05% by weight), high in protein(preferably from about 55 to about 60% by weight), finely milled(preferably 100% of the cottonseed flour is less than 75 micron in size)and contains carbohydrates at about 25 to 30%, preferably about 28% byweight. Additional carbohydrate sources may be included such as cane orbeet sugar. Likewise, additional lipid oil sources such as refinedcottonseed oil, corn oil, soybean oil, and the like, preferablycottonseed oil, may be added.

Fire ants may vary their preferences from day to day by variouslyseeking sugars, starches, proteins and oils for their diet. Therefore,an arrestant and feeding stimulant containing these four items would bea universal feeding stimulant satisfying the varying dietary demands ofthe fire ants.

The arrestant and feeding stimulant may have additional componentscomprising the various types of aids known to those skilled in the artwhich include but are not limited to diluents such as water, clays andthe like, formulation aids such as emulsifiers, application aids such asdispersants, wetting agents, spreader-stickers, and the like, and UVprotectors.

The arrestant and feeding stimulant is preferably used at rate per acreranging from 0.25 to about 10 lbs. based on the weight of the protein,carbohydrate and lipid oil sources. Preferably, the range is from about0.5 to about 2 lbs. per acre based on the weight of the protein,carbohydrates and lipid oil sources.

The pheromone in the biopesticide composition is optional in that thearrestant and feeding stimulant in and of itself is a food lure thatacts as an attractant to the targeted insect pests.

In general, pheromones or attractants may be classified as sex, food, oroviposition lures. Additional classifications or subclassificationsinclude trail pheromones, aggregating and other pheromones. Broadlydefined, a sex pheromone is an odor released by one member of thespecies which attracts the opposite member for the purpose of mating.The presence of sex pheromones has been demonstrated in most orders ofinsects and they can be produced by the male or female of the species.In many eases, it is the female which produces the attractant. A largenumber of pheromones that are useful in the biopesticide compositions ofthe present invention may be naturally or synthetically obtained.Pheromones and attractants are well known by those skilled in the art.Examples of such pheromones and attractants are disclosed in U.S. Pat.Nos. 4,908,977 and 3,895,078, the disclosures of which are incorporatedherein by reference. When included, the pheromone may be incorporated inthe biopesticide composition of the present invention in a controlledrelease form.

A preferred pheromone is the sex pheromone of the male boll weevil alsoknown as grandlure which is comprised of four monoterpenoid componentswhich are disclosed in U.S. Pat. No. 3,895,078. The components are (I)(+)-cis-2-isopropenyl-1-methylcyclobutaneethanol; (II)(Z)-3,3-di-methyl-delta¹,beta -eyclohexaneethanol; (III)(Z)-3,3-dimethyl-delta¹,alpha -cyclohexaneacetaldehyde; and (IV)(E)-3,3-dimethyl-delta¹,alpha -cyclohexaneacetaldehyde. These componentsare preferably present in the following range: Component I: from about30 to about 35% by weight, component II: from about 35 to about 40% byweight, component III: from about 13 to about 15%, and IV: from about 13to about 15% by weight. A particularly preferred ratio is about30:40:15:15, respectively. The rate of usage of the pheromone ispreferably from about 25 to about 1,000 milligrams per acre, morepreferably from about 40 to about 400 milligrams per acre.

In regards to boll weevils, the biopesticide composition is preferablyapplied when the outside ambient temperatures range from about 10° C. toabout 40° C. which is conducive to adult boll weevil flight.Applications are preferably made on a multiple basis at five (5) dayintervals until the population of boll weevils is below an economicthreshold. Application of the fungus and/or the biopesticide compositionmay be accomplished using standard operating equipment used in theagricultural industry, for example, a tractor or airplane equipped withbooms and nozzles for spraying the composition in a liquid carrier.

The following examples are for illustrative purposes only and are notmeant to limit the claimed invention in any manner.

EXAMPLES EXAMPLE No. 1 Feeding Substrate Evaluations

The boll weevil primarily feeds and reproduces on cotton although otherplants have been identified as alternative hosts. In the presentexample, commercially available materials containing cotton plant deriveproducts that were evaluated as potential feeding substrates for theadult boll weevil. The products evaluated were: Proflo which is acottonseed derived protein from Traders Protein, Memphis, Tex.; Konsume™from Fermone Corporation, Inc., Phoenix, Ariz.; and cottonseed oil(twice refined) from Valley Co-op Oil Mill, Harlingen, Tex. Konsume™feeding stimulant contains a mixture of cottonseed flour, disaccharide,vegetable lipid oil, alcohol alkoxylate (emulsifier), and polysaccharide(thickener).

Laboratory evaluations in this example and the following example wereperformed with boll weevils obtained from Robert T. Gast RearingLaboratory at Mississippi State, Miss., and reared by the method ofRobinson and Wright disclosed in Advances and Challenges and InsectRearing, pages 188-192, U.S. Department of Agriculture, Washington, D.C.(1984; editors E. G. King and N. C. Leppa).

Solutions of each of the potential feeding substrates in water wereprepared at concentrations of 0, 10, 30, 50, 80 and 100 percent or eachof the substrate materials. Five squares each about 400 to 600milligrams were immersed into each solution and placed in 100 millimeterglass petri dishes with ten boll weevils. The weevils were 3-5 days ofage, of mixed sexes and had not received food nor water for 24 hours.The number of adults observed feeding at time intervals of 30 and 60minutes and the number of feeding punctures in the squares at 24 hourswere the criteria used to differentiate between the substrates (10replications). Further the evaluations were made by placement of therespective materials on dental cotton rolls to determine if adults wouldfeed on the substrates if placed on an abstract material.

As noted in Table 1, adults fed readily on each of the three substratesevaluated. At the higher concentrations (30 percent plus) of allmaterials evaluated, fewer adults were observed to feed as thesubstrates were oily or sticky and apparently the physical conditionacted as a deterrent. Some mortality was observed at 24 hourpost-feeding for the cottonseed oil and Prorio substrates whereas nonewere observed with Konsume™ substrate. The number of feeding punctures(Table 1) provided data that supported the number of adults observed tofeed on the substrate treated squares. The Konsume™ substrate, in bothcases, was the choice as determined statistically with more adultsobserved to feed as well as more feeding punctures in the squares.

The next evaluation was to determine if adults would feed on substratesif placed on an inert material other than the fresh cotton squares.Cotton dental wicks were immersed in 10 percent solutions of cottonseedoil, Proflo and Konsume™, and the data in Table 2 show that the adultsfeeding on the substrates were not deterred by the placement of thesubstrates on the cotton dental rolls. No significant differences wereobserved among the three feeding substrates in feeding activity. Asnoted in Table 3, a comparison with treated squares and dental cottonroll reveal no significant differences when a choice was available tothe adults.

                  TABLE 1                                                         ______________________________________                                        Substrate evaluation for a biopesticide as determined                         by attractancy and feeding behavior of the boll weevil.                       Feeding Activity of Adults                                                                x ± se    x ± se                                            Substrate   observed feeding.sup.A                                                                     punctures/squares.sup.A                              ______________________________________                                        Cottonseed oil                                                                            2.11 ± 0.19 a                                                                           18.95 ± 2.56 a                                    Konsume ™                                                                              3.28 ± 0.21 b                                                                           29.52 ± 3.08 b                                    Proflo      2.89 ± 0.21 a                                                                           21.33 ± 3.06 a                                    ______________________________________                                         .sup.A Means followed by the same letter in each test are not                 significantly different, ttest analysis (P < 0.05).                      

                  TABLE 2                                                         ______________________________________                                        Substrate evaluation for a biopesticide as determined                         by attractancy and feeding behavior of adult boll weevil                      when the Substrate was placed on an inert material.                           Feeding activity of adults                                                    Substrate    x ± se observed feeding.sup.A                                 ______________________________________                                        Cottonseed oil                                                                             6.25 ± 4.25 a                                                 Konsume ™ 5.5 ± 4.5 a                                                   Proflo        8.0 ± 2.91 a                                                 ______________________________________                                         .sup.A Means followed by the same letter are not significantly different,     ttest analysis (P < 0.05).                                               

                  TABLE 3                                                         ______________________________________                                        Feeding behavior of adult boll weevils when Substrate                         was placed on squares or cotton wicks: choice evaluation.                     Feeding activity of adults                                                    Substrate on x ± se observed feeding.sup.A                                 ______________________________________                                        Squares      3.11 ± 0.27 a                                                 Cotton filters                                                                              2.8 ± 0.24 a                                                 ______________________________________                                         .sup.A Means followed by the same letter are not significantly different,     ttest analysis (P < 0.05).                                               

EXAMPLE 2 Laboratory Evaluation of an Embodiment of the Biopesticide

In this example, the combination of boll weevil pheromone (0.1milligrams per milliliter), feeding substrates (10%), andentomopathogenic fungus (1%) on dental rolls was placed on top of screencages holding adult boll weevils. The fungus was Beauveria bassiana inthe form of a technical powder obtained from Abbott Laboratories, NorthChicago, Ill.

Adult boll weevils were observed to readily feed on the combination andafter five days more than 90 percent were dead. Fungus growth wasexternally apparent. The pheromone was a grandlure preparation. The deadadults were dissected to determine the presence of Beauveria bassiana.

EXAMPLE 3 Preliminary Field Evaluation of Biopesticide

In this example, a preliminary field evaluation of the biopesticideinvolved the placement of ten milliliters of the solutions used in theprevious example on sanitary napkin pads located on stands two metershigh at the edges of a cotton field in the Fall of 1987. Numbers ofadults that responded to the biopesticide were visually counted aftertwo hours.

No difference in attractancy or feeding by adults was observed among thethree feeding substrates. It was difficult to quantify this type ofevaluation other than to record a number of adults at a given time toindicate that adults would or would not respond to the biopesticide. Theobservations provided data that indicated a positive response.

EXAMPLE 4 Overwintered Adults in Early Season Cotton

In this example, the biopesticide was evaluated against over-winteredboll weevil adults on cotton. The evaluation was initiated in the Springmonths on a 14-acre cotton block in North East Hidalgo County. The fieldwas divided into two-seven acre sections: one for the boll weevilbiopesticide and one for conventional insecticide treatments for earlyseason boll weevils. Each section was further divided into one acresubsections in each treatment. Pheromone traps that contained nopheromone were placed five per subsection (one acre) and monitored priorto planting of cotton and throughout the test. The objective of thisevaluation was to determine if adults were induced to come into thecotton early in response to the boll weevil biopesticide and todetermine if the fungus Beauveria bassiana (Abbott Labs.) would be aneffective pathogen.

At the six leaf stage of growth in the cotton, applications of the bollweevil bioinsecticide were initiated. The biopesticide contained about681 grams of conidia (one gram had about 1×10¹⁰ spores as correctedspore viability); about 8.5 liters of Konsume™; about 2.0 gramspheromone (grandlure preparation) in a total volume of about 170 liters.A highboy sprayer with 8,002 nozzles, six rowband at about 40centimeters high was used to apply about 19.4 liters per hectare. Adultboll weevils were placed in individual screened cages on the groundprior to the application of the bispesticide and removed immediatelythereafter to the laboratory for determination of pathogenicity ofBeauveria bassiana (25 adults per container with one container per acretreated subsection). After application, boll weevil adults (5 per planton 5 plants in subsection plots) were retained on the plants by 20centimeter plexiglass tubing for 24 hours and then removed to thelaboratory for fungal evaluation in the last two applications. Agarplates containing Sabouraud Dextrose Agar and Yeast Extract [about 40gm. dextrose, about 10 gms Neopeptone (Difco), about 15 gm. agar andabout 10 gm. yeast extract per 1000 mi. water] medium (See Fenge et al.,infra.) (14 in treated and 14 in non-treated subsample plots) werereturned to the laboratory and held at 95% relative humidity and 29° C.for determination of fungal growth.

Adults were removed from the non-baited pheromone traps from the treatedand non-treated plots and held in the laboratory to determine thepresence of the fungus. Five traps per acre gave a total of 70 pheromonetraps within the field.

As the cotton matures and few feeding and ovipositional sites areavailable near the termination of the cotton production season, adultboll weevils begin to search for additional food and over-winteringhabitat. The objective of the testing of the boll weevil biopesticide atthis time would be to attract those dispersing adults to thebiopesticide, on non-cotton habitat as well as regrowth cotton, anddetermining pathogenicity of the fungus. The regrowth cotton occurs whenthe crop is shreaded and not immediately plowed. The regrowth gives theadult boll weevils feeding sites and probably increases a number of theadults that survived the winter. Plowdown or the destruction of thecotton stocks has been a recommended and accepted agricultural methodfor reduction of overwintered adult population since the early 1900's.

The results of the biopesticide applications on adult boll weevils whenapplied to early season cotton are given in Table 4. Adults that wereplaced in the screened cages at the time of application will return tothe laboratory with the results that 78.02%±6.12% adults (number ofadults=641) were killed by the fungus Beauveria bassiana (Abbott Labs.).It is noted that these cages were placed at ground surface between thecotton plants and therefore the exposure to the Beauveria bassiana inthe biopesticide would have been less than total. Adult boll weevilsthat were caged on plants after application of the bioinsecticide had amortality factor of 92.54%±6.5% (number of boll weevils=139) due toBeauveria bassiana (see Table 4). Adult boll weevils captured in thenon-baited pheromone traps averaged 11% mortality due to Beauveriabassiana (number of boll weevils=100).

                  TABLE 4                                                         ______________________________________                                        Results of biopesticide applications on adult boll                            weevils when applied to early season cotton.                                  Pathogenic Activity of Beauveria                                              Method of Exposure                                                                            x % ± se mortality                                         ______________________________________                                        Screen cages    78.02 ± 6.12                                               Caged plants    92.54 ± 6.5                                                ______________________________________                                    

EXAMPLE 5 Non-Cotton Habitat Test

In this example, an area of rangeland grasses was identified as anon-cotton/boll weevil habitat and used to evaluate the attractancy ofthe boll weevil biopesticide during the period of adult dispersal. Plotsize was about 0.25 acre and three were treated with the boll weevilbiopesticide and three served as controls. The plots were three milesapart. Pheromone traps (5 unbaited) were placed equal distantly withinthe plots for evaluation of adults that were attracted by thebioinseetieide. Sweep nets were used to capture adults within the grass.The adults were returned and challenged in the laboratory for thepresence of the Beauveria bassiana (Abbott Labs.) which would indicatethat contact had been made with the bioinseetieide, whether per os orexternally. The boll weevil biopesticide formulation used was about 3.78liters of Konsume™ feeding substrate, about 1.89 liters of Nufilm 17,about 40 grams of calco oil red, about 400 milligrams of pheromone,about 100 grams of Beauveria bassiana and about 151.4 liters of water.Nufilm 17 is about 96% di-1-p-menthene, available from Miller Chemicaland Fertilizer Corp., Hanover, Pa. The Nufilm 17 acts as a stickerspreader, extender and antitransparent. The Beauvecia bassiana is of thestrain identified as RS-252 available from Abbott Laboratories, Chicago,Ill. Four applications at 4-day intervals were applied.

The results of the application of the boll weevil biopesticide torangeland grass plots indicated that a significant number of adult bollweevils were attracted to the treated plots when compared to nontreatedcontrols (P=<0.05, number=92). Of those adults captured either bynon-baited pheromone traps or by sweep nets, about were found to beinfected by Beauveria bassiena.

EXAMPLE 6 Regrowth Cotton Test

In this example, cotton was shreaded and allowed to regrow on eight (8)1-acre sites to provide an area in which to evaluate the boll weevilbiopesticide. Three applications at five-day intervals were made on theregrowth cotton and adult boll weevils were hand collected by visibleinspections of the plant or by the use of sweep nets. Four plots, about1 acre each, were treated and four were left untreated. Unbaitedpheromone traps, five per acre, also were placed on each plot. The bollweevil biopesticide and application was the same as used in Example 5 inthe non-cotton habitat treatments. Adults removed from the plots werereturned to laboratory and challenged for fungal activity.

The regrowth application of the bioinsecticide to the regrowth cottonplots resulted in about 60.1% infection with Beauveria bassiana inadults captured (number=158) in the treated plots.

EXAMPLE 7 Bait Stations

In this example, bait stations were developed using one gallon plasticmilk containers by removing the sides and placing a wire platform in thebottom so that about 300 milliliters of solution could be placed in thebottom and the adult boll weevils could land on the wire and feed. Thistest was evaluated in a brushy habitat at least 50 miles north of anycotton production. The bait stations, located on top of 2 meter stakes,were about one mile apart and the formulations evaluated were replicatedthree times with nine evaluations made. Adult boll weevils were removedand counted after an hour interval to determine attractancy. Adult bollweevils were returned to the laboratory (number=10 if available or less)and challenged for mortality. The Beauveria bassiana (Abbott Labs.) waspresent in the solution at a concentration of 1% (1×10¹⁰ conidia pergram). The feeding substrate, Konsume™, was diluted 50% with water andthree formulations with pheromone (10 milligrams per bait station) wereevaluated: an encapsulated pheromone; a formulated lure strip availablefrom Hercon Laboratories, South Plainfield, N.J.; and a grandlurepreparation.

As shown in Table 5, the evaluation of different pheromone formulationsin the bait station reveals significant differences in the number ofadults captured. The grandlure preparation when combined with thefeeding substrate and Beauveria bassiana attracted significantly moreadult boll weevils than the encapsulated pheromone or the lure strip.When the adult boll weevils were challenged for the presence ofBeauveria bassiana, no significant differences occurred between thosecaptured from the bait stations with the lure strip and the grandlurebut there was a significant difference between those caught and thesetwo the encapsulated pheromone/feeding substrate combination, as shownin Table 6.

                  TABLE 5                                                         ______________________________________                                        Evaluation of pheromone formulations in the                                   biopesticide placed in bait stations.                                         Numbers of adults captured in 1 hour                                          Formulation 10 mg..sup.a/                                                                      x ± se adults.sup.b/                                      ______________________________________                                        Konsume ™ ls  33.29 ± 6.93 a                                            Konsume ™ gl  64.92 ± 19.74 b                                           Konsume ™ ec   0.7 ± 0.24 c                                             Konsume ™ (alone)                                                                           0 d                                                          ______________________________________                                         .sup.a/ ls = lure strip gl = grandlure formulation; ec = starch               encapsulation; Konsume ™ = feeding substrate.                              .sup.b/ Means followed by the same letter are not significantly different     ttest (P < 0.05).                                                        

                  TABLE 6                                                         ______________________________________                                        Evaluation of pathogenicity of the                                            biopesticide when placed in bait stations.                                    Percent of adults infected with Beauveria                                     Formulations.sup.a/                                                                            x ± se Mortality.sup.b/                                   ______________________________________                                        Konsume ™ ls  37.77 ± 6.82  a                                           Konsume ™ gl   51.1 ± 22.64 b                                           Konsume ™ ec  14.7 ± 7.56 c                                             Konsume ™ (alone)                                                                           0 d                                                          ______________________________________                                         .sup.a/ ls = lure strip gl = grandlure formulation; ec = starch               encapsulation; Konsume ™ = feeding substrate.                              .sup.b/ Means followed by the same letter are not significantly different     ttest (P < 0.05).                                                        

EXAMPLE 8 Experimental Field Trials

Experimental field trials were conducted at the Subtropical AgriculturalResearch Laboratory in the lower Rio Grande Valley where the boll weevilis the major pest of cotton. The field experimental design was areplicated block, five times in the first trial, and three times in thesecond trial. Each plot was about 1 acre. Cotton cultivar was Gossypiumhirsutum L., "DES-119", on Irrigated Sandy Clay Loam Land. Plots in thefirst field trial were treated with Composition A and plots in thesecond field trial were treated with Composition B at approximately the8-leaf stage and then on a weekly schedule thereafter for a total of sixapplications. Following these applications with the biopesticide,conventional insecticides were then used throughout the remainder of thecotton production season. The application equipment utilized was a JohnDeere, 6000 Ground Rig Sprayer which delivered about 8 gallons per acre.

The treatments evaluated in the first field trial were Composition Aaccording to the present which utilized a Beauveria bassiana from AbbottLaboratories (strain RS-252) which was compared to control C1 which wasthe insecticide Bidtin only. Bidrin is(E)-2-dimethylcarbamoyl-1-methylvinyl dimethyl phosphate (CAS: 41-66-2).In the second field trial, the treatments were with Composition Baccording to the present invention, control C2 and control C3.Composition B utilized the fungus Beauveria bassiana, deposit ATCC-74040(also accessioned as ASREF-3097). Control C2 utilized the insecticideGuthion only and control C3 utilized the insecticide Guthion inconjunction with the feeding stimulant utilized in Composition B.Guthion is O,O-dimethyl-S-[(4-oxo-1,2,3-benzotriazin-3(4H)-yl) methyl]phosphoro dithioate (CAS: 86-50-0).

The formulations for compositions A and B are shown in Table 7. Otherthan the particular fungus used, the compositions were otherwise thesame. These were added to water to make a total volume of about 8gallons.

                  TABLE 7                                                         ______________________________________                                        Fungus:                                                                       Composition A      Beauveria bassiana                                                            (Abbott Labs.).sup.a                                       Composition B      Beauveria bassiana                                                            (ATCC-74040).sup.b                                         Arrestant/Feeding Stimulant:                                                  Formulation:.sup.c 1. cottonseed flour.sup.d                                                     2. cottonseed oil                                                             3. sugar (disaccharide)                                                       4. emulsifier.sup.e                                                           5. thickener.sup.f                                                            6. water and other inerts.sup.i                            Pheromone:         Grandlure.sup.g                                            Aids               Nufilm 17.sup.h                                            ______________________________________                                         .sup.a suspended in water with 0.05% Tween X100, a wetting agent; total       volume of the suspension was about 270 milliliters.                           .sup.b suspended in oil; total volume of the suspension was about 270         millliliters; fungus also accessiones as ARSEF 3097 by the USDAARS.           .sup.c Components 1-5 of the formulation compose about 34% by weight of       the formulation with component 6 composing the remainder. The protein,        carbohydrate and lipid oil content of components 1-3 combined is in a         weight ratio of about 42.5:55:2.5 of protein to carbohydrate to lipid oil     Total volume of the formulation was about 1 quart.                            .sup.d Defatted (<4% fat), low gossypol (<0.05%), high protein (55-60%),      finely milled (100% <75 microns) cottonseed flour containing about 28%        carbohydrates.                                                                .sup.e Arnox 2404, an alcohol alkoxylate available from Witco Corp.,          Houston, Tx.                                                                  .sup.f Xantham gum.                                                           .sup.g The weight ratio of components I, II, III and IV was about             30:40:15:15 respectively. Total weight of the Grandlure was about 400 mg.     .sup.h About 1 pint total volume.                                             .sup.i The other inerts in the formulation do not add or detract to the       arresting and feed stimulating properties of the formulation.            

The application rates were Bidrin (one pint per acre), Guthion (aboutone pint per acre), Beauveria Bassiana from Abbott Laboratories at4.54×10¹² conidia per acre and in Composition B Beauveria bassianadeposit ATCC-74040, at 6.26×10¹² conidia per acre.

Adult boll weevils in screened petri dishes were placed within the rowsat the time of application, then returned to the laboratory andchallenged for Beauveria bassiana (25 adults boll weevils per petridish: five dishes per plot per application). Adult boll weevils werealso placed on treated plants enclosed in plexiglass tubes, left for 24hours, then removed to the laboratory and challenged for Beauveriabassiana. (Five adults per plant: Five plants per plot per application).

Plant density, plant height, and fruit numbers (squares, bolls) clean,damaged, and total were taken at five sites within the plot. At each ofthese sites, measurements were taken at random for two one-meter rowareas at various times during early crop development to determine theimpact of early treatments with the biopesticide of the presentinvention.

Boll weevil pheromone traps (four) baited with 10 milligram lure stripsfrom Hereon Laboratories were located at the corners of the test field.During the four-month period of field trials 1 and 2, 722 and 517 adultboll weevils were captured, respectively. The results for field trials 1and 2 are shown in Tables 8 and 9, respectively. In each of the twofield trials, the biopesticide of the present invention yieldedsignificantly more than the control without treatment (Nontreated) andless than that of the insecticide treatment controls (C1, C2, and C3).The activity of the biopesticide is slower than that of the insecticidecontrols and as such the differences therebetween were expected. Theaddition of the feeding stimulant to the insecticide Guthion and theresults thereof substantiated the relative attractiveness of the feedingstimulant to adult boll weevils.

The mortality of the boll weevil adult exposed in petri dishes is givenin Table 10 and no significant differences were detected between the twofield trials in this regard. The pathogenicity of the Beauveria bassianais not immediate as that of a toxicant such as Guthion or Bidtin;however, the combined effect of contact activity of the fungus andingestion thereof within the feeding stimulant gives excellent mortalityfor a biopesticide. The mortality of adult boll weevils caged on plantsafter treatment is given in Table 11 and is at a similar level whencompared to Table 10. Also, activity is present for 72 hourspost-treatment in this evaluation method, although declining. (See Table11).

Plant height characteristics for field trials one and two, that is,plant height, plants for row meter, and fruiting levels are given inTables 12, 13, 14 and 15. The data indicate no difference isattributable to the treatments are present in any of the measuredparameters with the exception of fruit which is given in Table 14 whichat the time of measurement the non-treated plots had less than thetreated plots which were indicative of the boll weevil activity.

Table 16, status of cotton, provides the data that is reflected in thefinal yield data for the treatments and shows the effects of the earlyseason treatment using the biopesticide of the present invention as wellas that of the insecticide controls on the potential crop. Relativelevels of damage can be compared to show the effects of the biopesticideto that of the insecticide.

The change from a wettable powder formulation in Composition A to thatof a flowable suspension in Composition B improved considerably theapplication of the biopesticide. Composition B should also have improvedthe activity in that cuticles are lipophilie and are more susceptible tothe absorption of an oil whereas water-based formulations tend to runoff. The conidia of the fungus are hydrophobic and are not easily wettedso that the oil provides a better base for expression of contactactivity. An improvement in lint yield was noted from the first fieldtrial to the second field trial.

Statistical analysis of data in the present example was accomplishedwith an analysis of variance (ANOVA) and Tukeys HSD test (T-test) wasused to assign significant differences between individual treatments andthe non-treated control. (According to the procedure given in MStat.1987. Michigan State University). Unless indicated otherwise, P<0.05 wasthe criterion used for determining significant treatment effects.

                  TABLE 8                                                         ______________________________________                                        Mean lbs ± S.D. of lint production                                         per acre, Field Trial 1                                                       Treatment      Yield ± S.D..sup.1                                          ______________________________________                                        Control C1     367.0 ± 72.0 a                                              Composition A   216.9 ± 128.1 b                                            Nontreated     126.9 ± 47.9 c                                              ______________________________________                                         .sup.1 One way analysis of variance; means separated by Tukeys HSD test (     < 0.05). One acre plots with 5 randomized replications.                  

                  TABLE 9                                                         ______________________________________                                        Mean lbs ± S.D. of lint production                                         per acre, Field Trial 2.                                                      Treatment      Yield ± S.D..sup.1                                          ______________________________________                                        Control C2     431.1 ± 117.0 a                                             Composition B  281.0 ± 107.1 b                                             Control C3     482.2 ± 154.5 c                                             Nontreated     134.6 ± 50.5  d                                             ______________________________________                                         .sup.1 One way analysis of variance; means separated by Tukeys HSD test (     < 0.05). One acre plots replicated 3 times.                              

                  TABLE 10                                                        ______________________________________                                        Mortality of adults exposed in petri dishes after application                 Field Trial    N      % Dead ± S.D..sup.1                                  ______________________________________                                        1              1950   83.5 ± 17.5 a                                        2              2477   89.6 ± 9.3  a                                        ______________________________________                                         .sup.1 Means followed by same letter are not significantly different          (TTest, P < 0.05).                                                       

It will be apparent from the foregoing that many other variations andmodifications being made in the methods and the compositionshereinbefore described by those having experience in this technology,without departing from the concept of the present invention.Accordingly, it should be clearly understood that the methods andcompositions referred to herein in the foregoing description areillustrative only and are not intended to have any limitations on thescope of the invention.

                  TABLE 11                                                        ______________________________________                                        Mortality of adults caged on individual plants after treatment                Exposure                                                                      Post-Treatment                                                                           Field Trial % Dead ± S.D..sup.1                                 H          1       2       1989     1990                                      ______________________________________                                        24         510     375     81.0 ± 18.9                                                                         82.6 ± 15.5                            48         --      377     --       79.5 ± 23.6                            72         --      288     --       73.1 ± 12.5                            ______________________________________                                         .sup.1 Means followed by same letter are not significantly different          (TTest, P < 0.05).                                                       

                  TABLE 12                                                        ______________________________________                                        Plant characteristics, Field Trial 1.                                                   Plant Ht Plants/    Fruit/                                                    (cm) ± S.D.                                                                         M ± S.D.                                                                              M ± S.D.                                     ______________________________________                                        Composition A                                                                             34.8 ± 7.3 a                                                                          19.2 ± 7.1  a                                                                         118.6 ± 56.2 a                           Control C1  31.9 ± 4.7 a                                                                          20.1 ± 11.1 a                                                                          84.0 ± 35.1 b                           ______________________________________                                         .sup.1 Means followed by a different letter are significantly different       (TTest P < 0.05).                                                        

                  TABLE 13                                                        ______________________________________                                        Plant Height (cm) ± S.D. Field Trial 2.                                           Date of Sample                                                         Treatment                                                                              4/16    4/23    4/30  5/08  5/25  5/20                               ______________________________________                                        Control C3                                                                             14.0 ±                                                                             11.5 ±                                                                             17.5 ±                                                                           22.8 ±                                                                           41.9 ±                                                                           57.0 ±                                   2.1     2.2     2.0   3.0   6.5   4.0                                Control C2                                                                              9.6 ±                                                                             11.8 ±                                                                             17.9 ±                                                                           22.0 ±                                                                           42.5 ±                                                                           55.5 ±                                   2.3     2.3     3.1   4.4   8.0   8.8                                Composi- 10.4 ±                                                                             11.4 ±                                                                             17.6 ±                                                                           21.09 ±                                                                          35.6 ±                                                                           40.0 ±                          tion B   1.5     2.6     3.2   4.9   5.9   6.9                                Nontreated                                                                    ______________________________________                                    

                  TABLE 14                                                        ______________________________________                                        Fruit per meter row ± S.D., Field Trial 2.                                               Date of Sample                                                  Treatment       5/8      5/25                                                 ______________________________________                                        Control C3      2.6 ± 1.9                                                                           70.8 ± 18.5                                       Control C2      2.4 ± 1.6                                                                           89.3 ± 32.5                                       Composition B   2.2 ± 1.6                                                                           78.2 ± 26.9                                       Nontreated               51.6 ± 9.9                                        ______________________________________                                    

                  TABLE 15                                                        ______________________________________                                        Plants per meter row ± S.D., Field Trial 2.                                         Date of Sample                                                       Treatment  4/16    4/23     4/30  5/08  5/25                                  ______________________________________                                        Control C3 14.1 ±                                                                             12.6 ±                                                                              13.1 ±                                                                           16.1 ±                                                                           13.4 ±                                        3.9     3.9      2.7   3.3   3.6                                   Control C2 12.3 ±                                                                             12.1 ±                                                                              10.8 ±                                                                           15.7 ±                                                                           13.9 ±                                        4.1     3.4      3.0   5.0   6.7                                   Composition B                                                                            13.6 ±                                                                             11.7 ±                                                                              12.9 ±                                                                           14.3 ±                                                                           13.3 ±                                        4.6     3.9      2.9   3.2   2.5                                   Nontreated                              15.4 ±                                                                     3.1                                   ______________________________________                                    

                  TABLE 16                                                        ______________________________________                                        Status of cotton, Field Trial 2.sup.A.                                        Treatment                                                                     Control C2    Control C3                                                                              Composit. B                                                                              Nontreated                                 ______________________________________                                        Square/M                                                                      Clean   29.6 ± 27.0                                                                          26.1 ± 15.8                                                                          27.4 ± 24.6                                                                         0.8 ± 1.7                             Damaged 11.8 ± 11.8                                                                          19.4 ± 12.8                                                                          19.4 ± 17.7                                                                         1.3 ± 2.9                             Total   41.5 ± 34.1                                                                          41.9 ± 16.4                                                                          46.8 ± 38.0                                                                         2.1 ± 4.3                             Bolls/M                                                                       Clean   38.8 ± 18.6                                                                          34.3 ± 12.8                                                                          20.2 ± 11.1                                                                         5.1 ± 4.5                             Damaged 5.8 ± 5.3                                                                            9.1 ± 8.3                                                                            8.0 ± 5.8                                                                           17.5 ± 6.8                            Total   42.6 ± 18.0                                                                          41.9 ± 16.5                                                                          28.2 ± 12.9                                                                         22.6 ± 9.5                            Fruit/M                                                                       Damaged 17.6 ± 12.1                                                                          27.8 ± 16.9                                                                          27.5 ± 16.8                                                                         18.8 ± 7.9                            Total   84.5 ± 32.5                                                                          87.2 ± 26.7                                                                          74.9 ± 37.4                                                                         24.6 ± 10.7                           Plants/M                                                                              11.7 ± 3.8                                                                           12.1 ± 3.9                                                                           11.7 ± 3.3                                                                          15.7 ± 4.2                            ______________________________________                                         .sup.A M = meter row.                                                    

EXAMPLE 9 Bioassays of Beauveria Bassiana, Deposit ATCC-74040(ASREF-3097)

In this example, the cotton fleahopper, Psuedatomoscellis seriatus, andsweet potato whitefly, Bemisia tabaei nymphs and adults were dipped insolutions of the biopesticide Composition B (having about 1×10¹⁰ conidiaper milliliter) and observed for mortality due to pathogenicity of thefungus Beauveria bassiana, deposit ATCC-74040 (ARSEF-3097).

These laboratory bioassays against the cotton fleahopper and the sweetpotato whitefly suggest that both are receptive to the pathogenicity ofthis particular strain of Beauveria bassiana.

What is claimed is:
 1. A biologically pure culture of Beauveria bassianahaving the identifying characteristics of Beauvecia bassiana ATCC 74040.2. A biopesticidal composition comprising an effective amount of afungus having the identifying characteristics of Beauvecia bassiana ATCC74040 and an agriculturally acceptable carrier.
 3. The composition ofclaim 2 wherein said fungus is in the form of spores.
 4. The compositionof claim 2 wherein the carrier is in the form of a liquid, a powder,granules or small particles.
 5. The composition of claim 2 wherein thecarrier is a liquid comprising water and a wetting agent.
 6. Thecomposition of claim 2 wherein the carrier comprises a cotton plantderivative.
 7. The composition of claim 3 wherein the carrier is aliquid carrier having 2×10⁸ to 2×10¹⁴ spores of Beauveria bassiana permilliliter of carrier.
 8. A biopesticidal composition for controlling atargeted pest comprising a fungus having the identifying characteristicsof Beauveria bassiana ATCC 74040 and an arrestant and feeding stimulantfor the targeted pest.
 9. The composition of claim 8 further comprisinga pheromone for the targeted pest.
 10. The composition of claim 8wherein said targeted pest is selected from the group consisting of bollweevil, cotton fleahopper, and sweet potato white fly.
 11. Thecomposition of claim 9 wherein the targeted pest is boll weevil and saidcomposition contains boll weevil grandlure.
 12. The composition of claim8 wherein the feeding stimulant is a cotton derived feeding stimulant.13. A process for controlling a pest selected from the group consistingof boll weevil, cotton fleahopper, and sweet potato white fly comprisingapplying the Beauveria bassiana of claim 1 to the pest, to foliage ofplants or to soil around plants.
 14. A process for controlling a pestselected from the group consisting of boll weevil, cotton fleahopper,and sweet potato white fly comprising applying the composition of claim2 to the pest, to foliage of plants or to soil around plants.
 15. Aprocess for controlling a pest selected from the group consisting ofboll weevil, cotton fleahopper, and sweet potato white fly comprisingapplying the composition of claim 8 to the pest, to foliage of plants orto soil around plants.